Mitochondria are vital organelles responsible for energy production and they undergo dynamic morphological changes influenced by the cell’s metabolic state. Mitochondrial membrane potential is an essential component in the process of energy storage in the cells. The membrane potential plays a crucial role in the functioning of mitochondria, and it can influence mitochondrial velocity through various mechanisms. Here we report correlation studies on the mitochondrial dynamics and mitochondrial membrane potential in mammalian cells using our home-built sMx-SPIM imaging system. The mitochondria are tracked manually by a self-developed algorithm to find the speed. The speed of mitochondria was determined by targeting mitochondria stained with MitoTracker Deep Red CMXRos dye and measurement of mitochondrial membrane potential was done by tetramethylrhodamine methyl ester (TMRM). Experimental studies were conducted on mammalian cells in alive conditions, and results are provided.
Light sheet fluorescence microscopy (LSFM) works on fluorescence imaging, employing target-specific fluorescent proteins to label regions of interest in a sample. In this way, analyzing and manipulating information from different targets in a specimen requires labeling the specimen using multiple fluorophores. Multispectral imaging helps identify several target molecules simultaneously at once, thereby retrieving information from the entire volume of the sample. Here we report multispectral imaging studies on mammalian cells using our in-house built multispectral LSFM imaging system. Experimental studies were conducted on mammalian cells in alive conditions, and results are provided.
Photoacoustic imaging (PAI) is now a very promising imaging technique that provides image with sufficient depth, good resolution, and optical contrast. A conventional PAI system is relatively expensive and mechanically bulky. The study demonstrated that MEMS PMUT achieves miniaturized ultrasound (US) sensor element (either single element or an array of elements) with superior performance in terms of power consumption, flexibility, broader bandwidth, and sensitivity. This implies that these technologically novel qualities hold promise for the use of PMUT as an acoustic sensor in PAI systems in place of the conventional piezoelectric bulk element-based spherical ultrasound (US) transducer. We report our study on the design and development of MEMS PMUT−(central frequency ~ 1MHz) based PAM−that integrates MEMS technology and imaging technology (specifically, photoacoustic imaging (PAI)). In this work, we present a temporal integration of the signals over a certain number (~20) of pulsed light-induced PA waves against the conventional technique to acquire a single 1D PA signals/data corresponding to one individual optical pulse−induced PA waves. This means to say that the enhancement of imaging performance with the use of PMUT acoustic sensor is associated with a reduction of obtainable temporal resolution, i.e., a trade-off exists. With the applied temporal integration method SNR has been improved ~ 20dB. The preliminary study demonstrates that the integration of PMUT in PA imaging modality holds promise as a future (imaging) technology both for biological studies and their applications.
The eyes function through the cooperation of different ocular components, and complications with any components would lead to ocular diseases that deteriorate vision. Hence early and precise detection and monitoring of ocular diseases, along with the improved understanding of pathological mechanisms, becomes essential for successful treatment. Photoacoustic Imaging (PAI) is a non-invasive and non-destructive imaging modality based on the photoacoustic effect, which gives high spatial resolution, sensitivity, contrast and penetration depth. Since PAI can provide anatomic and functional ocular characterizations, it can be a potential tool for medical screening/diagnosis of ocular diseases, staging, treatment, and continuous post-treatment monitoring. Here, we perform deep-tissue imaging of the Goldfish (Carassius auratus) eye using a home-built Photoacoustic microscope to identify various ocular components like iris, crystalline lens, retina, optic nerve and blood vessels. The study can be extended to observe changes in these structures under different ophthalmic disease conditions.
In the past two decades, High-Intensity Focused Ultrasound (HIFU) has been actively investigated for inducing tissue ablation for tumours, healing arteries and even skin tightening for inducing facelifts. Being a non-contact and non-invasive procedure, HIFU is an attractive alternative over other procedures like radio-frequency ablations and microwave ablations as these are semi-invasive methods. Computer simulations in this domain can provide us with a fast-track way to test and deduce results virtually before conducting related experiments. In this study, we simulated the properties of bovine liver tissue virtually in a MATLAB-based k-Wave toolbox to track lesion variations with varying HIFU on-time and with a constant amount of heat deposition. Photoacoustic reconstructions were also carried out with a linear and circular array of transducers using the simple Delay-And-Sum (DAS) algorithm to access the real-time imaging capability of this technique for HIFU therapy. All the results obtained are discussed and validated. This study will, in general, aid in deciding the HIFU configuration and operational parameters with respect to the tissue type, lesion size, and lesion location within the tissue.
The improper disposal of textile dyes like Rhodamine B into nearby water bodies and land areas affects living organisms nearby. Here, we explore the effects of rhodamine absorption through the soil into the plant root and how variation in rhodamine concentration affects the growth and development of the plant. The developmental defects over time are verified by nuclear staining the plant cells. Furthermore, we hypothesis that the chemotropic effect seen in plants guides the root away from rhodamine B high concentration regions to low concentration regions. This will in turn help the plant recover from the injury. These minute structural variations are analyzed with the help of our in-house developed dual-arm multi-level magnification light-sheet microscopy (DMx-LSFM) system. The microscope consists of an automated (translating and rotating) sample holding stage, which helps to observe nearby regions by bringing the region of interest into a plane of focus without disturbing the sample.
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